Click here to close
Hello! We notice that you are using Internet Explorer, which is not supported by Xenbase and may cause the site to display incorrectly.
We suggest using a current version of Chrome,
FireFox, or Safari.
Biochem Biophys Res Commun
2014 Nov 14;4542:308-12. doi: 10.1016/j.bbrc.2014.10.070.
Show Gene links
Show Anatomy links
Mechanisms of Rose Bengal inhibition on SecA ATPase and ion channel activities.
Hsieh YH
,
Huang YJ
,
Jin JS
,
Yu L
,
Yang H
,
Jiang C
,
Wang B
,
Tai PC
.
???displayArticle.abstract???
SecA is an essential protein possessing ATPase activity in bacterial protein translocation for which Rose Bengal (RB) is the first reported sub-micromolar inhibitor in ATPase activity and protein translocation. Here, we examined the mechanisms of inhibition on various forms of SecA ATPase by conventional enzymatic assays, and by monitoring the SecA-dependent channel activity in the semi-physiological system in cells. We build on the previous observation that SecA with liposomes form active protein-conducting channels in the oocytes. Such ion channel activity is enhanced by purified Escherichia coli SecYEG-SecDF·YajC liposome complexes. Inhibition by RB could be monitored, providing correlation of in vitro activity and intact cell functionality. In this work, we found the intrinsic SecA ATPase is inhibited by RB competitively at low ATP concentration, and non-competitively at high ATP concentrations while the translocation ATPase with precursors and SecYEG is inhibited non-competitively by RB. The Inhibition by RB on SecA channel activity in the oocytes with exogenous ATP-Mg(2+), mimicking translocation ATPase activity, is also non-competitive. The non-competitive inhibition on channel activity has also been observed with SecA from other bacteria which otherwise would be difficult to examine without the cognate precursors and membranes.
Cabelli,
SecA protein is required for secretory protein translocation into E. coli membrane vesicles.
1988, Pubmed
Cabelli,
SecA protein is required for secretory protein translocation into E. coli membrane vesicles.
1988,
Pubmed
Cabelli,
Characterization of membrane-associated and soluble states of SecA protein from wild-type and SecA51(TS) mutant strains of Escherichia coli.
1991,
Pubmed
Chen,
Identification and characterization of protease-resistant SecA fragments: secA has two membrane-integral forms.
1998,
Pubmed
Chen,
The first low microM SecA inhibitors.
2010,
Pubmed
Chen,
ATP is essential for protein translocation into Escherichia coli membrane vesicles.
1985,
Pubmed
Chen,
A significant fraction of functional SecA is permanently embedded in the membrane. SecA cycling on and off the membrane is not essential during protein translocation.
1996,
Pubmed
Cui,
Design, synthesis and biological evaluation of rose bengal analogues as SecA inhibitors.
2013,
Pubmed
Driessen,
Protein translocation across the bacterial cytoplasmic membrane.
2008,
Pubmed
Economou,
SecA membrane cycling at SecYEG is driven by distinct ATP binding and hydrolysis events and is regulated by SecD and SecF.
1995,
Pubmed
Hsieh,
SecA alone can promote protein translocation and ion channel activity: SecYEG increases efficiency and signal peptide specificity.
2011,
Pubmed
,
Xenbase
Hsieh,
Reconstitution of functionally efficient SecA-dependent protein-conducting channels: transformation of low-affinity SecA-liposome channels to high-affinity SecA-SecYEG-SecDF·YajC channels.
2013,
Pubmed
,
Xenbase
Huang,
Fluorescein analogues inhibit SecA ATPase: the first sub-micromolar inhibitor of bacterial protein translocation.
2012,
Pubmed
Karamanou,
A molecular switch in SecA protein couples ATP hydrolysis to protein translocation.
1999,
Pubmed
Li,
Discovery of the first SecA inhibitors using structure-based virtual screening.
2008,
Pubmed
Lill,
The ATPase activity of SecA is regulated by acidic phospholipids, SecY, and the leader and mature domains of precursor proteins.
1990,
Pubmed
Lin,
Electrophysiological studies in Xenopus oocytes for the opening of Escherichia coli SecA-dependent protein-conducting channels.
2006,
Pubmed
,
Xenbase
Lin,
Escherichia coli membranes depleted of SecYEG elicit SecA-dependent ion-channel activity but lose signal peptide specificity.
2012,
Pubmed
,
Xenbase
Linnertz,
Erythrosin 5'-isothiocyanate labels Cys549 as part of the low-affinity ATP binding site of Na+/K+-ATPase.
1998,
Pubmed
Linnertz,
Molecular distance measurements reveal an (alpha beta)2 dimeric structure of Na+/K+-ATPase. High affinity ATP binding site and K+-activated phosphatase reside on different alpha-subunits.
1998,
Pubmed
Manting,
SecYEG assembles into a tetramer to form the active protein translocation channel.
2000,
Pubmed
Mao,
Maximal efficiency of coupling between ATP hydrolysis and translocation of polypeptides mediated by SecB requires two protomers of SecA.
2009,
Pubmed
Matsuyama,
SecD is involved in the release of translocated secretory proteins from the cytoplasmic membrane of Escherichia coli.
1993,
Pubmed
McNicholas,
SecA proteins of Bacillus subtilis and Escherichia coli possess homologous amino-terminal ATP-binding domains regulating integration into the plasma membrane.
1995,
Pubmed
Mitchell,
Two distinct ATP-binding domains are needed to promote protein export by Escherichia coli SecA ATPase.
1993,
Pubmed
Nakatogawa,
Two independent mechanisms down-regulate the intrinsic SecA ATPase activity.
2000,
Pubmed
Oliver,
Azide-resistant mutants of Escherichia coli alter the SecA protein, an azide-sensitive component of the protein export machinery.
1990,
Pubmed
Pogliano,
SecD and SecF facilitate protein export in Escherichia coli.
1994,
Pubmed
Ramamurthy,
Topology of the integral membrane form of Escherichia coli SecA protein reveals multiple periplasmically exposed regions and modulation by ATP binding.
1997,
Pubmed
Rao C V,
Antibiotic targeting of the bacterial secretory pathway.
2014,
Pubmed
Segers,
Traffic jam at the bacterial sec translocase: targeting the SecA nanomotor by small-molecule inhibitors.
2011,
Pubmed
Tai,
In vitro protein translocation into Escherichia coli inverted membrane vesicles.
1991,
Pubmed
Tanfani,
Effects of fluorescent pseudo-ATP and ATP-metal analogs on secondary structure of Na(+)/K(+)-ATPase.
2000,
Pubmed
Wang,
Ring-like pore structures of SecA: implication for bacterial protein-conducting channels.
2003,
Pubmed
Wickner,
Escherichia coli preprotein translocase.
1996,
Pubmed
You,
Phospholipids induce conformational changes of SecA to form membrane-specific domains: AFM structures and implication on protein-conducting channels.
2013,
Pubmed
Yu,
Expression, purification, and characterization of Pseudomonas aeruginosa SecA.
2006,
Pubmed
Zhang,
Specificity of SecYEG for PhoA precursors and SecA homologs on SecA protein-conducting channels.
2013,
Pubmed
,
Xenbase
van der Wolk,
The low-affinity ATP binding site of the Escherichia coli SecA dimer is localized at the subunit interface.
1997,
Pubmed
